LOCUS       sequence-199761-        7002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses synaptophysin fused jRGECO1a under the synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7002)
  TITLE     AAV plasmids to package compartmentalised calcium indicators
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7002)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-synp-F-synaptophysin-jRGECO1a
FEATURES             Location/Qualifiers
     source          1..7002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1721..1730
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1742..1798
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     2494..2513
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    2987..3020
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3071..3104
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3129..3717
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3182..3202)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3600..3611)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3749..4225
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3948..3967)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4265..4405
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4265..4394
                     /label=AAV2 ITR
     rep_origin      4480..4935
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4567..4586)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4777..4798
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4952..4971)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5071..5093
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5131..5149)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5217..5321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5322..5390,5391..6182)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5540..5559)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6353..6941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6842..6861
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgccgcacc atggacgtgg tgaatcagct ggtggctggg ggtcagttcc gggtggtcaa
      841 ggagcccctt ggcttcgtga aggtgctgca gtgggtcttt gccatcttcg cctttgctac
      901 gtgtggcagc tacaccgggg agcttcggct gagcgtggag tgtgccaaca agacggagag
      961 tgccctcaac atcgaagttg aattcgagta ccccttcagg ctgcaccaag tgtactttga
     1021 tgcaccctcc tgcgtcaaag ggggcactac caagatcttc ctggttgggg actactcctc
     1081 gtcggctgaa ttctttgtca ccgtggctgt gtttgccttc ctctactcca tgggggccct
     1141 ggccacctac atcttcctgc agaacaagta ccgagagaac aacaaagggc ctatgatgga
     1201 ctttctggct acagccgtgt tcgctttcat gtggctagtt agttcatcag cctgggccaa
     1261 aggcctgtcc gatgtgaaga tggccacgga cccagagaac attatcaagg agatgcccat
     1321 gtgccgccag acagggaaca catgcaagga actgagggac cctgtgactt caggactcaa
     1381 cacctcagtg gtgtttggct tcctgaacct ggtgctctgg gttggcaact tatggttcgt
     1441 gttcaaggag acaggctggg cagccccatt catgcgcgca cctccaggcg ccccggaaaa
     1501 gcaaccagca cctggcgatg cctacggcga tgcgggctac gggcagggcc ccggaggcta
     1561 tgggccccag gactcctacg ggcctcaggg tggttatcaa cccgattacg ggcagccagc
     1621 cagcggtggc ggtggctacg ggcctcaggg cgactatggg cagcaaggct atggccaaca
     1681 gggtgcgccc acctccttct ccaatcagga tccaccggtc gccaccatgg tcgactcatc
     1741 acgtcgtaag tggaataagg caggtcacgc agtcagagct ataggtcggc tgagctcacc
     1801 cgtggtttcc gagcggatgt accccgagga cggcgccctg aagagcgaga tcaagaaggg
     1861 gctgaggctg aaggacggcg gccactacgc cgccgaggtc aagaccacct acaaggccaa
     1921 gaagcccgtg cagctgcccg gcgcctacat cgtggacatc aagttggaca tcgtgtccca
     1981 caacgaggac tacaccatcg tggaacagtg cgaacgcgcc gagggccgcc actccaccgg
     2041 cggcatggac gagctgtaca agggaggtac aggcgggagt ctggtgagca agggcgagga
     2101 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     2161 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg aggcctttca
     2221 gaccgctaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     2281 ccctcagttc atgtacggct ccaaggccta cattaagcac ccagccgaca tccccgacta
     2341 cttcaagctg tccttccccg agggcttcag gtgggagcgc gtgatgaact tcgaggacgg
     2401 cggcattatt cacgtgaacc aggactcctc cctgcaggac ggcgtattca tctacaaggt
     2461 gaagctgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg
     2521 ctgggaggct acgcgtgacg acctgactga agagcagatc gcagaattta aagaggcttt
     2581 ctccctattt gacaaggacg gggatgggac gataacaacc aaggagcttg ggacggtgtt
     2641 ccgatcgctg gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga
     2701 tgccgacggt gacggcacat tcgacttccc tgagttcctg acgatgatgg caagaaaaat
     2761 gaatgacaca gacagtgaag aggaaattcg cgaagcgttc cgcgtgtttg ataaggacgg
     2821 caatggctac atcggcgcag cagagcttcg ccacgtgatg acagaccttg gagagaagtt
     2881 aacagatgag gaggttgatg aaatgatcag ggtagcagac atcgatgggg atggtcaggt
     2941 aaactacgaa gagtttgtac aaatgatgac agcgaagtag gctagcataa cttcgtataa
     3001 agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat tctttagaat
     3061 ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc gatatcaagc
     3121 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3181 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3241 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3301 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3361 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3421 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3481 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3541 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3601 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3661 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3721 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     3781 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     3841 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     3901 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     3961 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     4021 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     4081 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     4141 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     4201 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     4261 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     4321 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     4381 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     4441 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4501 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     4561 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4621 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4681 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4741 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4801 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4861 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4921 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4981 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5041 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5101 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5161 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5221 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5281 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5341 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5401 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5461 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5521 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5581 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5641 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5701 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5761 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5821 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5881 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5941 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6001 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6061 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6181 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6241 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6301 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6361 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6421 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6481 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6541 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6601 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6661 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6721 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6781 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6841 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6901 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6961 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
